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Experiments were performed on K562 cells engineered by piggyBac transposition of fusion protein constructs (except opAsCas12a was delivered by lentiviral transduction). Flow cytometry analysis of CD81 expression knockdown 6 days after transduction of the indicated lentiviral crRNA constructs. Shown are averages of median single-cell expression knockdown from two to five biological replicates for each crRNA construct, with error bars indicating standard error of the mean. One-sided Wilcoxon rank-sum test was performed for differences in single-cell expression distributions ( n > 200 cells) for each fusion protein against multiAsCas12a-KRAB for each replicate. Asterisk indicates P < 0.01 for all replicates for a given pairwise comparison.

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